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A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects

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Schlipalius, D. I., Waldron, J., Carroll, B. J., Collins, P. J. and Ebert, P. R. (2001) A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects. Insect Molecular Biology, 10 (6). pp. 579-585. ISSN 1365-2583

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Article Link: https://doi.org/10.1046/j.0962-1075.2001.00297.x


Existing procedures for the generation of polymorphic DNA markers are not optimal for insect studies in which the organisms are often tiny and background molecular information is often non-existent. We have used a new high throughput DNA marker generation protocol called randomly amplified DNA fingerprints (RAF) to analyse the genetic variability in three separate strains of the stored grain pest, Rhyzopertha dominica. This protocol is quick, robust and reliable even though it requires minimal sample preparation, minute amounts of DNA and no prior molecular analysis of the organism. Arbitrarily selected oligonucleotide primers routinely produced ~50 scoreable polymorphic DNA markers, between individuals of three independent field isolates of R. dominica. Multivariate cluster analysis using forty-nine arbitrarily selected polymorphisms generated from a single primer reliably separated individuals into three clades corresponding to their geographical origin. The resulting clades were quite distinct, with an average genetic difference of 37.5 ± 6.0% between clades and of 21.0 ± 7.1% between individuals within clades. As a prelude to future gene mapping efforts, we have also assessed the performance of RAF under conditions commonly used in gene mapping. In this analysis, fingerprints from pooled DNA samples accurately and reproducibly reflected RAF profiles obtained from individual DNA samples that had been combined to create the bulked samples.

Item Type:Article
Subjects:Science > Entomology
Science > Biology > Genetics
Live Archive:09 Jan 2024 23:33
Last Modified:09 Jan 2024 23:33

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